Arizona State University-Tempe Campus

NIH Research Projects · FY 2025 · 2022-09

Project Summary More than 6 million U.S. adults live with diagnoses of Alzheimer's Disease and Related Dementias (AD/ADRD), with projections as high as 13.8 million cases by 2050. There are no cures for AD/ADRD, making lifestyle factors key targets for reducing risk, as they account for at least a third of AD/ADRD cases. Engaging in regular physical activity (PA), particularly in midlife, is associated with reduced risk for AD/ADRD. Yet nearly half of midlife adults (48%) do not meet national PA guidelines of 150 minutes/week of moderate-intensity PA. Goal setting is a commonly used behavior change strategy to increase PA. Key psychosocial mechanisms believed to underpin the use of goal setting to promote PA include self-regulation and self-efficacy. Yet, the most effective goal setting technique to enhance these psychosocial mechanisms for the successful promotion of PA and adherence to national PA guidelines remains unclear. In the proposed study, we will use a two- phased approach to empirically test three goal setting techniques to enhance psychosocial mechanisms of self- regulation and self-efficacy for the successful promotion of PA and adherence to national PA guidelines among insufficiently active midlife adults with obesity. In the R61 phase, a Phase 1 pilot study will establish feasibility and help refine the intervention. In the R33 phase, a Phase 2 9-month 4-arm proof-of-concept mechanistic trial (6-month active intervention and 3-month no contact follow-up) will be implemented to establish preliminary efficacy of goal setting techniques to increase PA and promote adherence to national PA guidelines. All participants will receive a Fitbit to self-monitor PA and engage in PA action planning sessions with a study interventionist. In addition, participants will be randomly assigned to 1 of 4 groups: i) static weekly goal of 150 minutes/week of moderate-intensity PA, which most closely resembles the approach of public health campaigns and care providers; ii) weekly self-selected PA goals, which allows for self-determination and adaptation of the goal; iii) modest incremental weekly PA increase goal (i.e., researcher determined PA goal that 20% minutes/week greater than the minutes/week of PA in the previous week); or iv) non-goal setting control group. Based on Goal Setting Theory, it is hypothesized that participants in the incremental goal group will have the greatest increases in self-regulation and self-efficacy, which in turn, will lead to the greatest improvements in PA and adherence to national PA guidelines over the 9-month intervention. Further, individuals with the greatest PA adherence are expected to experience the greatest improvements in AD/ADRD risk factors, including cognitive functioning (memory, executive functioning, processing speed), AD plasma biomarkers (plasma amyloid 42/40 ratio. plasma phosphorylated tau 231), and measures of cardiometabolic disease risk (BMI, blood pressure, plasma glucose, TG, total cholesterol, LDL-C, HDL-C, VLDL-C, IDL-C, insulin and insulin sensitivity).

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY/ABSTRACT Genomics science gives us a better understanding of the interactions between genes and the environment, thus allowing researchers to find better ways to improve health and prevent diseases. The Training in Genomics Research (TiGeR) Track will engage students from under-represented populations, such as those from racial and/or ethnic minority groups, people with disabilities, those from sexual and gender minority groups, and those from disadvantaged backgrounds. The program will target qualified candidates from these under-represented communities with undergraduate degrees in computer science, mathematics, or statistics with limited or no experience working with genomics datasets. The TiGeR Track program’s main goal is to prepare students for successful careers as computational experts in genomics research. This goal will be accomplished through our targeted recruitment and retention efforts within the two- year graduate curriculum culminating in a master’s degree in biological data science with a concentration on genomics research. The TiGeR Track will be integrated into our recently established Master’s in Biological Data Science (MSBDS) degree program. The MSBDS is an interdisciplinary program that capitalizes on the three pillars of specialty within the School of Mathematical and Natural Sciences (SMNS) at Arizona State University (ASU) West: computer science, life sciences, and mathematics. However, the MSBDS currently has no focus on genomics research nor bioinformatics, and the TiGeR Track will help fill this gap. In addition to extensive course work and supervised hands-on research activities, the TiGeR students will be trained in AnVIL - a cloud-based platform for computational genomics as well as other genomics toolsets using publicly available datasets and repositories. Combining a strong focus on genomics research and large-scale data analytics skills will equip our TiGeR graduates to be immediate and valuable contributors to industry and research jobs in the genomics science sector. We will target, recruit and retain a diverse pool of students to develop into the next generation's thought leaders in genomics sciences. By recruiting students that represent the communities surrounding our institution, we will maximize deeper ties with surrounding communities at the regional level and generate a diverse science workforce that can compete globally. The TiGeR Track program will provide a model of interdisciplinary collaboration in genomics sciences among faculty within SMNS and other stakeholders on and off-campus. To be prepared to undertake a career in genomics science, the TiGeR Track graduates require a strong foundation of skilled creativity and the ability to identify and achieve scientific goals of significance to the biomedical and biosciences research enterprise. Throughout their tenure in the TiGeR Track program, the students will develop skills to utilize and analyze genomic datasets from the public domain in a research laboratory setting. While a robust interdisciplinary and immersive instruction structure is already present for data science training within the MSBDS degree program, additional resources are required to establish the genomics science curriculum, particularly in a way that will support students from under-represented backgrounds. Hence, we are submitting this grant application.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY Protein analysis is essential to the understanding of molecular scale processes in living systems, the diagnosis of diseases based on molecular biomarkers, and the treatment of diseases with drugs. The basic tasks of protein analysis include detecting a protein, identifying it and determining its interactions with other proteins or molecular ligands. Various technologies have been developed to perform these tasks, but the most indispensable ones are gel and capillary electrophoresis, Western Blot (WB) and enzyme linked immunosorbent assay (ELISA). These technologies separate and identify proteins based on a protein’s charge, size, and specific binding to antibodies. For molecular interaction analysis, surface plasmon resonance and other detection technologies are the current choices. Although ubiquitous in both research labs and industry, these platforms must be combined to provide complete analysis of proteins, which is complicated and time consuming. In addition, they lack single molecule analysis capability required for studying heterogenous processes and for achieving precision diagnosis, especially for low volume samples. The present project aims to develop one detection platform that can perform the key functions of the above technologies with single molecule detection capability. The proposed technology images single proteins without labels, measures the size, charge and mobility of each protein simultaneously, identifies the protein based on its specific binding to antibodies, and quantifies its interactions with other proteins in real time. The team at the Biodesign Center for Bioelectronics and Biosensors, ASU, has carried out substantial experiments to demonstrate the proposed technology. In this R01 project, the team will address remaining technical challenges, build a complete prototype and validate it for single protein analysis on single cells.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY / ABSTRACT Speech disorders are the most prevalent form of communication disorders. Many treatments of speech disorders involve learning new speech behaviors or modifying abnormal speech behaviors. These treatments strongly rely on processes of speech motor learning—improvement in motor performance through practice. However, speech disorders are often associated with impairments in various speech motor learning processes, resulting in inefficient or deficient speech motor learning. Inefficiencies in speech motor learning processes reduce the effectiveness of the treatments that rely on these learning processes. Therefore, there is a critical need (1) to understand the specific contributions of each of the processes of speech motor learning and (2) to develop behavioral protocols that selectively influence various motor learning processes to improve speech motor performance. Without this knowledge, the promise of developing effective and optimized treatments for speech disorders will likely remain limited. This proposal’s overall objective is to develop behavioral protocols that selectively target and improve speech motor learning processes in healthy adults. Here, we propose a research program to develop and optimize a set of visually augmented training protocols to improve the accuracy of two processes crucial for successful speech motor learning: auditory error detection and auditory-to-motor mapping. Our central hypothesis is that improving these processes through augmented training can improve speech motor learning. We formulated this hypothesis based on current theoretical models of speech, including our recent computational model. Aim 1 will evaluate the effects of error-detection training on speech motor learning. Using visual feedback and auditory feedback perturbation, we will train subjects to detect and estimate auditory errors more accurately. Subsequently, we will evaluate subjects’ extent of speech motor learning. Aim 2 will determine the contributions of auditory-to-motor mapping to speech motor learning. Using visual feedback, we will train subjects to learn the relationship between various articulatory configurations and their auditory consequences. Then, we will evaluate subjects’ extent of speech motor learning. This project’s results will have a critical positive impact because (1) they will form a strong scientific foundation that can inform the development of effective and optimized treatments for speech disorders, and (2) they will have significant theoretical implications by elucidating processes of speech motor learning.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY/ABSTRACT By 2060, 44% of older U.S. adults will be non-Hispanic White compared with 60% in 2020. Cognitive decline in this growing racially and ethnically diverse older population is a major public health concern. Blacks are disproportionately affected by Alzheimer’s disease and related dementias (ADRD), and potentially experience a faster rate of cognitive decline than other groups. Yet, little is known is about the mechanisms underlying this increased prevalence. Genetic mechanisms do not account entirely for the higher risk of decline and ADRD in older Black adults, suggesting that other factors may explain this variation. However, the majority of cognitive aging research derives from studies that have compared Blacks to Whites versus exploring within-group variability in health conditions, biological, and psychosocial risk and protective factors that may account for these differences. This study's overall goal is to clarify risk and protective factors underlying cognitive decline in Blacks by: (1) Examining the relation between high blood pressure (HBP) and domain-specific cognitive decline and exploring how serious life events and social support modify the HBP-cognition association; (2) Exploring the relation between inflammation and cognitive decline and how HBP mediates the relation between serious life events and cognition; and (3) Identifying the consequence of perceived stress on changes in cognition and evaluating whether HBP and social support moderates the stress-cognition relationship. The research plan will leverage valuable secondary data from two nationally recognized studies of older adults. This K01 Award application will facilitate the training and professional development of Dr. Byrd to launch her career as an independent investigator in the field of aging, cognition, and ADRD. The five-year training plan will fill gaps in (1) cognitive decline and aging research and (2) stress and coping theories, while building on her methodological strengths to include (3) advanced longitudinal methods. Dr. Byrd will complete the proposed research in the rich training environment of Arizona State University (ASU), with co-primary mentorship from David Coon, PhD, Professor and geropsychologist (ASU) and Roland J. Thorpe, PhD, Professor of Health, Behavior and Society and minority aging expert (John Hopkin’s University) and nationally and internationally recognized secondary mentors Peter Lichtenberg, PhD (geriatric neuropsychologist), Keith Whitfield, PhD (expert on cognition among Blacks) ,Toni Antonucci, PhD (social relations across the lifespan) and renowned biostatistician Wassim Tarraf, PhD. The proposal addresses Goals B and F of the NIA Strategic Directions for Research on Aging, which calls for research that seeks to understand 1) the effects of personal, interpersonal, and societal factors on aging and 2) health disparities among older adults. Our research has the potential to influence individual outcomes as well as affect policies and programs aimed at improving cognitive health for the aging population, particularly for Black Americans who are at greatest risk for cognitive decline and ADRD.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY/ ABSTRACT Enabled by the development of high-throughput, low-cost nucleic acids sequencing technologies, there have been accelerated development in genomics and transcriptomics in the past two decades, profoundly reshaping our knowledge in biology and medicine. However, similar technologies have yet to emerge for rapid identification and quantification of proteins. This is attributed to more complex structures of proteins, lack of polymerase chain reaction-like amplification methods, cellular heterogeneity, etc. Conventional protein sequencing methods, such as Edman degradation and mass spectrometry, are slow, expensive, and not suitable for detecting low- abundance proteins. Such ensemble measurements also can mask our fundamental understandings on how cells of a particular genotype function and respond to therapeutics. Single-molecule protein sequencing (SMPS) is an emerging research direction that directly reads amino acids sequence from individual protein or peptide molecules. Yet, a promising strategy using fluorosequencing still relies on long Edman degradation cycles and bulky fluorescent microscopes, not ideal for fast and low-cost readout. Electronic SMPS technologies using tunneling or nanopore sensors are emerging methods for development of portable and inexpensive sequencing systems. However, they still face challenges in precise nanofabrication, structural instability, high electronic noise, and inability in detection of all amino acids. To address the multi-faceted challenges in next-generation SMPS, we will design an on-chip integrated, electronic system that incorporates nano-opto-fluidic structures to transduce protein fingerprints into electronic signals at a high speed, a low cost and a small system foot-print. Our platform features an all-sapphire nanopore (AlSaPore) fluidic device that has a small capacitance and a greatly improved structural stability, and accordingly suited for high-speed, low-noise, high-throughput, electronic detection. Further, an ultrathin nitride-based metasurface-integrated circuit (MIC) structure is created on the AlSaPore to optically interrogate the fluorescently tagged single amino acids passing through the nanopore without conventional fluorescent microscopy. Subsequently, the fluorescent tag signals are transmitted through the waveguide and collected by on-chip integrated photodetectors. The optoelectronic channel (IA for amino acids tags) and ionic current channel (IP for protein primary structure) will be synchronously recorded, classified by deep learning algorithms, and used in combination to improve the protein sequencing accuracy. Supported by well-established nitride-on-sapphire device design and manufacturing technology, our MIC-AlSaPore is a scalable and compact platform that achieves single-molecule sensitivity with a potential to read out all 20 amino acids. The development of MIC-AlSaPore platform will have far-reaching impact in biomedicine beyond protein sequencing. It may be used for studying DNA-protein interactions at single-molecule levels, classification of specific genes, genome mapping and de novo assembly. Additionally, it may inspire future multi-omic (genomic, transcriptomic, and proteomic) diagnostic solutions with potential clinical applications.

NIH Research Projects · FY 2025 · 2022-09

Project Summary/Abstract The goal of the ASU Biomarker Characterization Center is to improve ovarian and lung cancer screening through the development of biologically-relevant circulating immune biomarkers. The scientific approach of our Center is based on several fundamental principles. First, that altered cancer protein expression, structure, and post- translational modifications induce host autoantibodies to create circulating biomarkers. Second, that alterations in microbial antigen expression (such as respiratory pathogens) also induce immunity, often detected in benign rather than malignant disease. Third, that the protein modifications, as well as the immune response to these neoantigenic structures, are heterogeneous between people, and that serologic biomarkers may complement circulating protein biomarkers. We will take a systems immunology approach to discover three types of antibodies, anti-microbial antibodies, autoantibodies and anti-aberrant glycoprotein antibodies. Our proposal builds on our extensive experiences with cancer biomarker discovery and immunoproteomics technology development. Our previous results on autoantibody biomarkers have been confirmed in blinded phase 2 multicenter validation studies and led to a CLIA-certified commercial blood test. Our results have shown that multiplexed panels of autoantibodies are required for adequate predictive value. With prior EDRN support, we have developed a set of innovative immunoproteomics technologies, namely high-density nucleic acid programmable protein array (HD-NAPPA), contra-capture protein array (CCPA) and multiplexed in solution protein array (MISPA), that, together with the largest full-length human and microbial gene collection at our DNASU plasmid repository, enable us to study antibodies against the full human proteome, microbial proteomes and the human O-glycoproteome for antibody biomarker signatures in cancer. Our Meso Scale Diagnostics (MSD) team has fielded over 3,000 instruments worldwide, and over 700 commercially available biomarker assay kits. Our expertise at serologic assay development was selected by Operation Warp Speed to use the V-PLEX® serology panels as the basis of its standard binding assays for immunogenicity assessments in all funded Phase III clinical trials of COVID vaccines. We will use our MSD MultiArray platform to migrate the top serologic and protein markers for their utility in our target clinical applications. We will collaborate with experts on lung and ovarian cancer screening at Vanderbilt University Medical Center, Boston University, MD Anderson Cancer Center, and German Cancer Research Center, who will also provide access to high-quality well-characterized samples to develop circulating biomarkers to enhance ovarian cancer screening or to distinguish benign from malignant pulmonary nodules. Adhering to the principles of PRoBE design, we will perform Phase I discovery by screening protein arrays with cancer patient and control sera for cancer or control-specific antibodies. Candidate biomarkers for both lung and ovarian cancers will undergo Phase 2 validation.

NIH Research Projects · FY 2026 · 2022-09

Project Summary/Abstract Large segments of current US middle-aged adults are suffering and doing more poorly than earlier-born cohorts43,65. These trends are distinct to the US. Several studies have documented that compared to Asian and European nations, US middle-aged adults exhibit higher rates of disability and chronic illness than same-aged peers6,10,73. Despite known differences between the US and other nations, little research has examined the mechanisms underlying diversity in midlife development and individual-level characteristics that either magnify or mitigate differences. Our objective is to use the HRS family of studies to identify the intergenerational, financial, and health behavior mechanisms driving diversity in midlife development and whether they differ across individual-level characteristics that span socio-demographics (i.e., education, gender, and race) and psychosocial resources (i.e., social support and control beliefs). Our specific aims are: (1) describe similarities and differences in midlife development of health, well-being, and cognition across multiple nations; (2) investigate the intergenerational, financial, and health behavior mechanisms driving diversity in midlife development; and (3) to examine the role of individual-level characteristics that moderate diversity in midlife development. To address our research questions, multilevel models will be applied to harmonized longitudinal data (collected from early 1990s to present day) from 17 nations across North America, Asia, and Europe. Each dataset contains national samples that are repeatedly assessed on interdisciplinary outcomes. The achievable outcome of Aim 1 is to comprehensively examine similarities and differences between the US trends and those obtained for other high-income nations. For Aim 2, we draw from the research team's conceptual and empirical research55,61 to investigate whether changing intergenerational relationships, increased financial vulnerabilities and a shrinking health behavior safety net are driving diversity in midlife development within- and between-nations. For Aim 3, we will target key questions of diversity by identifying population segments within and across nations who are most vulnerable and identify malleable protective resources to promote better positive midlife development. This project will provide novel insights into identifying the intergenerational, financial, and health behavior mechanisms underlying diversity in midlife development and whether they are operating similarly across nations with different policy contexts. For example, in nations with less comprehensive health care, such as the US, health behavior factors could be a driving force as opposed to nations with a national health care system. Similarly, we will uncover whether greater intergenerational dependency in the form of increased contact and financial transfers are a more potent mechanism in nations with less encompassing family and work policies. Findings from our project will also shed light on factors that can promote resilient outcomes and inform future prevention and intervention efforts.

NIH Research Projects · FY 2025 · 2022-09

Negative social experiences are a main driver of HIV disparities among Latino men who have sex with men (LMSM), negatively affecting non-injection substance use (e.g. methamphetamine) and PrEP uptake. Addressing factors that influence PrEP cascade progression, such as negative social experiences and methamphetamine use, are essential to ending the HIV epidemic. Negative social experiences can occur across various sources (i.e. family, friends, others), mechanisms (i.e. anticipated, enacted), and types, which compound to produce and exacerbate deleterious health outcomes among LMSM. Social network analysis techniques can be leveraged to examine how LMSM interact with and are affected by network members and inform targets for meaningful interventions focused on network members that (1) provoke negative social experiences and (2) provide social support. This K01 will provide the candidate, Dr. Algarin, with skills to conduct complex longitudinal multi-level structural equation modeling to examine the direct and indirect effects between negative social experiences and methamphetamine use risk and PrEP cascade progression through coping and resilience (AIM 1) and how sources of negative social experiences moderate these pathways (AIM 2) leveraging data from 500 LMSM enrolled in the NEXUS study (R01MH123282; PI: Smith). Using a community-engaged approach, Dr. Algarin will draw from these analyses to adapt a multi-level intervention to address negative social experiences through coping and resilience to decrease methamphetamine use risk and improve PrEP cascade progression (AIM 3). As an emerging scholar, Dr. Algarin will leverage the UCSD training infrastructure to build his professional independence and skills to promote HIV prevention interventions, positioning him as the next generation of scholars to end the HIV epidemic in the US. Specifically, Dr. Algarin seeks training to advance his statistical capacities (T1), gain proficiency multi-level intervention theories and concepts (T2) and multi-level intervention development and adaptation (T3), gain additional training in the ethical conduct of research (T4), and build skills for his professional development (T5). This K01 will provide Dr. Algarin with the necessary training and data to forge his research independence and produce a competitive future R34 to test the community- engaged, multi-level intervention he adapts as part of this study.

NIH Research Projects · FY 2024 · 2022-08

Project Summary: This proposal describes a five-year research and career development program to prepare Dr. Hamed Arami for a career as an independent investigator. This program will build upon Dr. Arami’s multidisciplinary background as a bioengineer scientist, trained in nanomedicine and basic cancer imaging, by providing expertise in brain cancer biology and image-guided therapy of brain tumors using Magnetic Particle Imaging (MPI). The PI will be mentored at Stanford Medical School by Drs. Sanjiv S. Gambhir (Main mentor, basic cancer biology, cancer pathology and cancer nanotechnology), Heike Daldrup-Link (co-mentor, magnetic nanomedicine, imaging and therapeutics), Max Wintermark (co-mentor, neuroimaging and brain MPI), Melanie Hayden (co-mentor, neurosurgery and neurology) and Bob Sinclair (co-mentor, nanomaterials characterization). Treatment of malignant primary brain tumors particularly glioblastoma multiforme (GBM) is challenging because of GBM resistant to chemotherapy and radiotherapy. Also, there are different types of GBM tumors that are not operable due to their locations in the brain (e.g. deep brain regions). In addition, routine GBM imaging in clinics are based on using gadolinium-based magnetic resonance imaging contrast agents. However, using these gadolinium-based contrast agents raises major concerns for GBM patients suffering from chronic kidney disease, which can be resolved by using nanoparticle contrast agents that do not show any renal clearance due to their larger size. The overall goal of the proposed research is to use MPI as a two-armed and high-resolution approach for safer imaging and magnetothermal therapy of the GBM. Four types of brain tumors with different levels of aggressiveness will be studied to identify the feasibility of the proposed method in different brain tumor microenvironments. Recently, I developed methods for tuning iron oxide nanoparticles (NPs) to generate high resolution (i.e. ~600 µm) MPI images with ultra-high contrast agent mass sensitivity of less than ~550pg Fe/µL. I have used MPI for three-dimensional targeted imaging of the U87 brain tumors in mice after intravenous injection of these NPs. Additionally, in separate studies, I demonstrated the feasibility of the MPI for selective magnetothermal therapy of the U87 tumors, when NPs were directly injected into tumors. In this project, I will first evaluate MPI and heat generation efficiency of the NPs at different brain depths to further identify ideal NPs design and imaging criteria for general brain tumor imaging or local magnetothermal therapy with MPI (Aim 1). Then, I will evaluate MPI for targeted 3D imaging of four different types of intracranially implanted brain tumors after intravenous injection of the nanoparticles, followed by nanoparticle biodistribution studies (Aim 2). Finally, I will use intratumoral injection of my tumor-penetrating NPs for MPI-guided magnetothermal therapy of the deep brain tumors (representative models for inoperable GBM), followed by in-depth survival and neuropathological studies (Aim 3). Iron oxide nanoparticles have been approved by FDA for several clinical applications and we hope that this method will ultimately find applications to many other types of solid tumors.

NIH Research Projects · FY 2024 · 2022-08

ABSTRACT Child maltreatment is associated with health risk behaviors carrying high personal and societal costs, such as substance use and sexual risk behavior.1–4 Placement of children into out-of-home care aims to mitigate these negative effects.5 Findings about rates of health risk behavior among youth in out-of-home care are mixed,6–16 although studies show that young adults formerly in care have heightened levels of health risk behavior.17–20 These mixed findings may be due to the fact that most studies test effects of placement across childhood on later health risk behaviors. However, placement during adolescence may be a turning point that redirects trajectories of health risk behavior. Adolescence is a period of rapid change and a typical period for initiation of these behaviors,21 and adolescents have more difficulties adjusting to placement than do younger children.22 To test whether adolescent placement acts as a turning point, it is important to know whether placement initiates long-term changes in trajectories, short-term (time-specific) changes, or both. To design interventions, it is also necessary to understand the mechanisms underlying placement effects. However, because placement may occur at various points across time, identifying its direct and mediated effects requires modeling out-of-home placement, mediators, confounders, and outcomes as time-varying variables. Importantly, out-of-home placement is a nonrandom event, and many pre-placement variables likely predict both placement and subsequent health risk behavior. To deal with this threat to causal inference, recent research has utilized propensity score methods.23–27 However, these methods cannot accommodate time- varying variables, precluding tests of placement as a turning point or tests of its time-specific mediators. Accordingly, the proposed study uses two modern methods for causal inference, inverse probability of treatment weighting (IPTW) and g-estimation, to test direct and mediated effects of placement on health risk behaviors across time. These methods can account for biases from observed confounders and accommodate time-varying variables.28,29 Aim 1 tests whether adolescents with out-of-home placement since the prior wave show short-term, time-specific increases in substance use and initiation of unprotected sex, compared to those who remain in-home. Aim 2 tests whether adolescent placement predicts longer-term elevations in adult health risk behavior. Aim 3 tests time-varying mediators of the effect of placement on adult health risk behavior. These aims will be tested in a national sample of 738 maltreated youth (age 11-14 at baseline) studied across four waves to adulthood. The training plan brings together a distinguished mentoring team to provide multidisciplinary training in child welfare system research, modern quantitative methods for causal inference, developmental psychopathology of health risk behaviors, implications for intervention and policy, and professional development. This will provide the applicant with the foundation to launch a research career studying the effects of placement and child welfare involvement on adolescents' developmental outcomes.

NIH Research Projects · FY 2025 · 2022-08

SUMMARY Metabolites generated by the gut microbiota provide multiple health benefits. However, the mechanism via which the gut microbiota utilizes substrates from the host and generates beneficial products are not well understood. Additionally, information regarding the role of secretome i.e., a collection of extracellular proteins secreted by the gut microbiota on health is minimal. Currently, no model system for the study of Type 9 protein Secretion System (T9SS) of gut microbes exists. Our preliminary bioinformatics analysis suggests the presence of functional T9SS in several human gut isolates of the genus Bacteroides. We predict that the T9SS of gut Bacteroides secretes pectate lyase and other enzymes that breakdown dietary fibers and subsequently ferment them to short- chain fatty acids (SCFAs). Reduction of SCFAs results in metabolic disorders. We predict that the relatively understudied, yet important, human gut isolates that include Bacteroides intestinalis, Bacteroides nordii, Bacteroides fluxus, Prevotella copri, and Parabacteroides distasonsis secrete many proteins via T9SS. We are developing a genetically tractable species B. intestinalis as a model organism to fill knowledge gap regarding studies of T9SS secretome in the gut. We hypothesize that the T9SS secretome of the gut microbiota might create a common pool of oligosaccharides that enrich SCFA producing species. Additionally, we aim to find controllers of a putative Bacteroides sensory transduction network that senses the abundance of dietary fibers and regulates production of SCFA producing enzymes. Our preliminary data also predicts the secretion of immune-suppressive cysteine proteases by the T9SS of B. intestinalis. The proposed experiments to test our predictions and hypothesis will significantly enhance our understanding of interspecies cooperation, resource optimization, and immunomodulation by the gut microbiota. T9SS is a recently discovered protein export pathway of bacteria of the Gram-negative Fibrobacteres-Chlorobi-Bacteroidetes superphylum. Thus far, the model organisms for the study of T9SS are from the human oral microbiota, environmental isolates, and pathogens infecting aquatic animals. With this proposal, we are using our expertise with T9SS to push the barriers that impede our understanding of secretome of the gut microbiota. The nuts and bolts of T9SS machinery are composed of nineteen different proteins but their structure and functional properties are unclear. In future, this information can help us control T9SS of the gut microbiota. At the core of T9SS is a rotary motor that powers gliding motility of microbes. This motility enables cargo transportation and shapes the spatial organization of a microbial community. We propose experiments to fill the gap regarding the mechanism via which T9SS enables bacterial motility and protein secretion. Two important knowledge gaps: (a) the role of T9SS in the gut microbiota, and (b) the structure and function of T9SS will be filled via a multi- pronged experimental approach that uses genetics, biochemistry, biophysics, and murine models. Together, we aim to gain a mechanistic understanding of T9SS machinery, its secretome in the gut microbiota, and their impact on health.

NIH Research Projects · FY 2025 · 2022-08

Project Summary Cell death has a critical role in human development and recovery following injury or disease. This is because dying cells produce signals that can significantly impact the behavior of the surrounding cells. The identity and consequences of these signals are diverse and context dependent, but many are known to regulate the survival, activity and proliferation of neighboring cells following injury. Thus, a better understanding of how dying cells impact surviving tissue could uncover novel therapeutic interventions to improve healing and regeneration following injury or disease. While this signaling phenomenon has been characterized in apoptotic cell death, it is unclear whether unregulated forms of cell death, such as necrosis, have a similar impact on tissue behavior and repair. Necrosis is the rapid, disordered death of cells, which can occur in any tissue and is central to many human conditions, including traumatic injuries (burns, frostbite), infections, and ischemic injuries like strokes and heart attacks. Several factors released from necrotic cells have been identified, however, the identity of other signals and whether they influence recovery has yet to be examined. The aim of this proposal is to investigate how necrotic wounds impact surrounding tissues to influence recovery and regeneration. Evidence that signals from dying cells impact nearby tissues first originated from studies of the larval wing primordia in Drosophila, called imaginal discs. These tissues have significant regenerative capacity, the study of which has led to important insights into the genetic events necessary for damage-induced tissue recovery. However, most of these studies examine apoptosis-induced regeneration, limiting our understanding of how cell death impacts surviving tissue to this type of injuries. To overcome this limitation, we have established a genetic tool that allows us to trigger either necrosis or apoptosis in the developing wing disc, and to genetically manipulate the surrounding cells that respond to each type of damage. With this tool we found that discs successfully regenerate in each case, but via different mechanisms. Notably, necrosis leads to widespread apoptotic cell death at a distance from the wound. This necrosis- induced apoptosis, or NiA, is necessary to drive regenerative proliferation and is therefore critical for proper recovery. The cause of NiA and how it promotes regeneration are currently unknown. Here, we propose to characterize the genetic response that leads to successful regeneration following necrosis focusing on the role of NiA. Our work aims to identifying how necrosis leads to NiA, understand how NiA promotes regeneration, and comprehensively characterize the necrosis-induced regeneration program that results in NiA using whole genome sequencing approaches. Together, the results of these experiments will contribute to our fundamental understanding of tissue repair in response to necrosis, which is ultimately essential for developing novel therapeutic approaches to treat necrotic wounds and promote regeneration in humans.

NIH Research Projects · FY 2024 · 2022-08

PROJECT SUMMARY (See instructions): Multimodal integration is a fundamental feature of animal nervous systems that allows them to extract useful information from complex environmental signals and guide behavior. A defect in this process may lead to communication disorders. However, there is currently limited understanding of this pervasive phenomenon. Due to its simplicity, the honeybee antennal lobe (AL) provides an excellent system for studying sensory integration. Chemical (i.e., odor) and mechanical (i.e., wind speed) information converge within the AL, likely in service of two intermingled tasks facing honeybees - tracking highly turbulent odor plumes and discriminating odors. Both are critical for foraging success. This proposal seeks to: (1) determine the impact of mechanical input (wind speed) on AL odor responses and odor classification; (2) determine the functional roles and mechanisms of multisensory integration within the AL. We postulate that the AL can switch between two distinct dynamic regimes – an odor tracking regime (triggered by high mechanical input) and an odor discrimination regime (triggered by low mechanical input). In other words, input from one modality affects the coding scheme of the other. To test this hypothesis, our experimental work will entail a suite of electrophysiological experiments that disentangle the contributions of each modality to AL dynamics, determine the impact of mechanical input on correlations across the AL, and assess the dependence of AL odor classification on mechanical input. Computationally, we will construct a realistic, experimentally benchmarked spiking network model of the AL integrating mechanical and olfactory inputs, and use it to study the network mechanisms that underlie AL dynamics within the two postulated regimes. The model will be used to explore conceptual ideas and generate specific hypotheses that will be tested in subsequent experiments. Finally, we will incorporate the fundamental principles uncovered in our work into novel machine learning algorithms for solving multimodal problems. The PIs are excellently suited for the proposed work – Dr. Lei is an expert in olfaction and the electrophysiological studies of the AL, Dr. Patel has extensive experience in biologically realistic modeling of AL dynamics, and Dr. Bazhenov is an expert in computational neuroscience, data analysis and machine learning.

NIH Research Projects · FY 2023 · 2022-08

Project Summary/Abstract Obstructive sleep apnea (OSA) is highly prevalent and associated with a spectrum of cardiovascular (CV) diseases and adverse health outcomes. However, OSA treatment strategies tend to show inconsistent treatment efficacy across individuals and little or no reduction in risk of CV diseases, events, or death. Phenotype discovery is critical for precise risk stratification and targeted treatment of OSA. Substantial heterogeneity among OSA patients is likely an important contributor to the suboptimal results of clinical trials. Thus, it is critical to delineate the OSA heterogeneity and stratify patients into high-vs low-risk clusters (i.e., “phenotypes”) associated with markedly different outcomes for precise risk stratification and targeted treatment. OSA data hold great promise to facilitate OSA phenotype discovery. Rigor of Prior Research: (1) We and others identified new prognostic factors in OSA data that are associated with one or more adverse CV outcomes. (2) Emerging OSA phenotypes were defined by machine learning and clustering algorithms from multi-faceted OSA data. (3) Newly identified OSA phenotypes, predictive of patients’ benefit from OSA treatments and risk for adverse CV outcomes, laid the foundation for OSA phenotypes’ clinical utility in targeted treatment and precise prognosis. However, significant gaps exist in fully leveraging the OSA data for phenotype discovery: There is a lack of “outcome-predictive”, “clinically-interpretable”, and “reproducible” phenotypes, defined from multi-domain OSA data in a large diverse U.S. population. To address these gaps, we propose a secondary multi-study analysis that seeks to develop new classification criteria and identify phenotypes in OSA by integrating multi-domain OSA-related sleep common data elements, including but not limited to patient socio-demographics, health habits, medical history, anthropometrics, polysomnography measures, daytime sleepiness, quality of life, and cardiovascular comorbidities and mortalities, combined across three of the largest epidemiological study cohorts deposited in the NIH-funded National Sleep Research Resource (NSRR). This includes Sleep Heart Health Study, Hispanic Community Health Study, and Multi-Ethnic Study of Atherosclerosis, with at least 5,336 OSA patients from a diverse population of African American, Caucasian, Hispanic, and Asian American men and women. Aim 1: Develop a novel sparse, outcome-predictive multi-domain Factor Mixture Model for OSA phenotype identification from multi-domain mixed-typed patient pre-clinical features and clinical features. Aim 2: Apply the developed model in Aim 1 to individual and pooled NSRR datasets to: (1) identify, characterize, and validate OSA phenotypes; (2) evaluate consistency and reproducibility in findings supported by individual and pooled analyses. Impact: We will identify, characterize, and validate OSA phenotypes that assist clinicians with determining how aggressive to be with the treatment plans and assist researchers with selecting appropriate patients to enroll in clinical trials of OSA treatment, eventually leading to precise prognosis and treatment of OSA.

NIH Research Projects · FY 2026 · 2022-06

Project Summary The goal of the proposed research is to understand the polymodal activation of TRPV1, specifically its activation by heat, protons, and chemical ligands. Understanding the molecular mechanisms that underlie TRPV1 function has significant implications in human health. TRPV1 is a polymodally regulated ion channel that is activated by many diverse stimuli, including heat, protons (low pH), and chemical ligands, like capsaicin, the pungent vanilloid from chili peppers. Over the past decade, there has been significant interest in developing TRPV1 antagonists to combat many types of pain and other relevant indications. One of the main complications in TRPV1 therapeutic intervention is that antagonists commonly dysregulate body temperature. Recent computational modeling and meta-analysis of human clinical trials suggest which modes of TRPV1 should be targeted for the development of analgesic antagonists that mitigate off-target effects. This proposal aims to dissect the independence, interdependence, and crosstalk between canonical TRPV1 activation modes and decipher the respective mechanisms. Nuclear magnetic resonance spectroscopy (NMR) and electrophysiology techniques will be used to achieve these goals. These data will be used to understand which TRPV1 regions underlie particular functions and illuminate allostery, cooperativity, and crosstalk between activation modes. To achieve these goals, two specific aims will be carried out. Aim 1 will focus on the characterization of a minimal TRPV1 construct inspired from natural TRPV1-isoforms that recapitulates the features of the full-length channel with electrophysiology and NMR studies. Additionally, this aim will provide the first structures of a human TRPV1 domain. A membrane domain that is responsible for ligand binding and involved in thermosensing. The structural studies will access non-cryogenic temperatures giving rise to information about the mechanism of thermosensing. These mechanistic and structural studies will be validated and contextualized with cellular studies. Aim 2 will focus on dissecting the allostery and crosstalk between TRPV1 heat, proton, and ligand modes of activation. One series of experiments will rely on validating computational predictions of human TRPV1 allosteric networks with whole- cell patch-clamp measurements. Another set of experiments will leverage chemical ligands from preclinical experiments and clinical trials that will be used with mutagenesis to identify ligand binding sites and how TRPV1 mode selectivity is achieved. The last sub-aim will employ an NMR-detected ligand screen of TRPV1 agonists and antagonists which will be subjected to emerging statistical analysis and learning techniques to generate methodologies capable of predicting which modes of activity TRPV1 modulators will activate. Significant preliminary electrophysiology and NMR data coupled with computational analysis indicate the feasibility of these aims during the timeframe of this proposal. The proposed biophysical and functional TRPV1 studies aim to better understand the molecular mechanisms that govern the function and complicate druggability and are anticipated to guide the development of the next generation of TRPV1 antagonists.

NIH Research Projects · FY 2026 · 2022-06

PROJECT SUMMARY Chronic low back pain (LBP) is one of the major contributors to musculoskeletal pain, disability, and lost workdays in the United States. LBP is highly correlated with degeneration of the intervertebral disc. Discogenic pain, specifically, is the leading cause of LBP. Patients with chronic discogenic LBP often exhibit aberrant sensory nerve growth deep within the disc. Once nerves are present in the degenerate disc they can be subjected to a complex milieu of pro-inflammatory mediators and dynamic mechanical loads causing stimulation and therefore pain. Binding of pro-inflammatory mediators to receptors on nociceptors results in lowered stimulation thresholds of ion channels responsible for action potential generation. Whereas mechanical loading of the disc can cause direct stimulation of mechanosensitive ion channels resulting in pain sensation. Previous work has explored the use of local delivery of an anti-inflammatory to the disc, however, incomplete pain resolution was observed. No work has examined targeting mechanoreceptors in the disc to alleviate chronic discogenic LBP. Further, no studies have yet been able to parse out which the role of nerve presence, inflammation, or mechanical loading is the major contributor to emergent pain. This question remains unanswered due to a lack of animal models that accurately mimic human presentation of chronic discogenic LBP. Taken together, these data support our overarching hypothesis that the presence of new nociceptors in the disc combined with inflammation and/or mechanical loading leads to chronic discogenic LBP, and targeted peripheral therapies may alleviate LBP. To test this hypothesis, we have developed an animal model of chronic discogenic LBP that exhibits a robust behavioral pain phenotype and nerve infiltration deep within the disc. By using innovative rodent models and targeted blockade of nerve growth, inflammatory mediators or mechanical sensing we will probe the roles of nerve presence, inflammation, and loading in LBP. We will then use this information to develop novel approaches to alleviate chronic discogenic LBP. Further, findings will be validated in aged animals which are more representative of a human LBP patient. The proposed work will use novel approaches using targeted neuro-inhibition, anti-inflammatories and mechanoreceptor antagonism to decouple the role of nerve presence, inflammation, and mechanical loading in LBP. The outcomes from this work include and better understanding of the specific mechanisms of LBP, laying the foundation to develop peripherally targeted pain for an intractable and widespread condition that is increasing in prevalence.

NIH Research Projects · FY 2026 · 2022-06

A large and rapidly growing majority of American Indian/Alaska Native (AI) families now reside in urban areas. Although they experience severe health disparities associated with substance abuse, risky sexual behavior, depression and suicide, few evidence-based prevention interventions address their distinctive needs. Family disruption, stresses due to migration and poverty, and cultural and social losses are often implicated in adverse health outcomes for urban AI families. By improving effective parenting skills and overall family functioning, culturally grounded parenting interventions enable parents to model and promote their children's well-being and reduce their children’s vulnerability to risk behaviors. The proposed study extends the project team's prior research on a culturally grounded parenting intervention for urban AIs, Parenting in 2 Worlds (P2W), which was co-developed with a coalition of urban Indian non-profit organizations, tested in three Arizona cities, and demonstrated efficacious. The research team joins two AI and two non-AI investigators, who together have extensive experience conducting collaborative research with AI populations in urban and tribal settings. This proposed multi-regional study will create new knowledge in four areas. First, the study will test P2W’s effectiveness beyond Arizona in improving parenting and family functioning, among a wider and more diverse group of urban AI communities located in cities spread across four regions: Northeast (Buffalo/Niagara), Midwest (St. Paul/Minneapolis), Mountain (Denver), and Southwest (Phoenix). Through the auspices of collaborating urban Indian center partners, the trial will recruit 720 families of AI youth age 12-17 (180 per city) and individually randomize them to receive P2W or an informational family health curriculum. Second, the study will test for moderators of the effectiveness of P2W, whether desired outcomes vary by the level of socioeconomic vulnerability, experiences of historical loss, or AI cultural identity of the parent participants. Third, the study will expand on the original Arizona trial to examine the adolescent’s reports on an enlarged range of youth health behaviors potentially impacted by the P2W intervention, including mental health (depressive symptoms, suicidality) as well as substance use and risky sexual behaviors. Fourth, the study will test for mediation—whether positive changes in parenting and family functioning that result from P2W lead to changes in the youth health behaviors. This would be the first cross-site multiregional trial of a culturally grounded parenting intervention designed specifically for urban AIs. It will advance critical knowledge on community prevention interventions for an under-served group severely affected by health disparities, and establish whether urban Indian centers and their communities can readily employ P2W to strengthen urban AI families and promote the behavioral health of their youth. It will also provide a foundation for advancing knowledge on effective prevention interventions in urban AI communities that have different migration histories and tribal compositions.

NIH Research Projects · FY 2025 · 2022-04

Abstract Our overarching goal is to use new quantitative methods of capturing dynamic family interactions in early childhood to identify key mechanisms underlying genetic and family intervention effects on substance use and dependence (SUD) and mental health problems across adolescence. Results will demonstrate the value of observational data analyzed with modern statistical approaches and inform the specification and refinement of more potent and effective family-based interventions. Our approach builds on findings from large sample genome wide association studies to inform formation of polygenic scores that represent genetic risk for SUD and differential susceptibility to both risky and promotive family relationship dynamics. Developmental theory on gene-environment interplay has highlighted the need to move away from focusing only on simple main effect models, thus, we examine genetic association in the context of dynamic social interactions and random assignment to a family-based intervention. Although SUD is heritable, it develops and progresses within problematic family interactions and relationships. Family interventions are central to evidence-based approaches to preventing and treating SUD, and direct observation of family interactions is the most rigorous way of measuring family interactions. To date, the methodology and analysis of family interactions relevant to intervening on youth SUD relies on broad aggregate scores. The most commonly used aggregate scores that define the family interaction may simply miss pathogenic dynamics. With the advancement of statistical analyses, there is unprecedented potential for accelerating observational family research over the coming decade. The proposed study involves secondary analysis of existing videotaped observations of racially/ethnically diverse children and families from the Early Steps Multisite Trial, applying dynamic structural equation modeling and multivariate multilevel survival analysis to understand the effects of polygenic risk and family intervention on downstream adolescent SUD and mental health problems. Early Step (N=731) is a randomized trial with long-term follow-up (ages 2-19) of the effects of the family-centered intervention, Family Check-Up, on reducing problem behaviors and SUD. The extensive data include videotaped observations of children and parents across multiple contexts at ages 2, 3, 4, and 5. In addition, children were genotyped using the contemporary Affymetrix Biobank Array. Such data provide a rare and unique opportunity to utilize new statistical methods to understand early habitual family dynamics on SUD and mental health problems in adolescence. These dynamic mechanisms provide key intervention targets for enhancing intervention effectiveness and efficiency and lead to enhancing potency of family-based interventions.

NIH Research Projects · FY 2026 · 2022-04

PROJECT SUMMARY/ABSTRACT American Indian and Alaska Natives (AIANs) experience disproportionately high rates of lifetime substance use disorder with peak past-year prevalence rates at 16 years of age. AIAN adolescents ages 17 and younger have a higher prevalence and earlier initiation of drug and alcohol use compared with all other ethnic/racial groups in the US. Compared to other ethnic/racial groups, AIAN youth have the highest self-reported depression rates, and in 2014, suicide was the second leading cause of death for AIANs between the ages 10 and 34 years of age. These data demonstrate the need for early intervention to prevent substance use in AIAN adolescents. Cultural connectedness has been identified as a protective factor against substance use and depression for AIAN youth while promoting positive self-esteem and healthy identity formation. Culturally- grounded after-school programs (ASPs) use cultural values and practices as a basis for health promotion and disease prevention, a form of AIAN prevention. Culturally-grounded program differ from other cultural programs (culturally-based or culturally-adapted) because the development is guided or led by the local community, for the local community and are rooted in specific social and cultural contexts. This study builds on a 6-year partnership with ASPs serving AI adolescents to assess the impact of a 13-session culturally-grounded intervention, called Native Spirit (NS). NS is an ASP that is led by traditional knowledge holders with each session focusing on a community-specific cultural value and activity. NS aims to decrease and prevent substance use by enhancing protective factors, including strengthening cultural identity, self-esteem, and resilience. The goals of the proposed (R00) study are to: 1) strengthen self-esteem, resilience, and cultural identity, and 2) attenuate substance use among AI youth (ages 12-17) through participation in NS. This study will use a mixed methods waitlist control design to evaluate the impact of participation in the NS program. The study will measure changes to participant self-esteem, cultural identity, resilience, and substance use with a self-report survey at three timepoints and semi-structured interviews. This study provides an innovative connection between cultural engagement and substance use prevention for AI adolescents and also highlights unique opportunities for health promotion with collaborations with ASPs that serve AI communities. Through the MOSAIC K99/R00, I will gain training in qualitative and quantitative analysis, culturally-appropriate measurement of adolescent substance use, and the development and implementation of prevention interventions while transitioning to an independent investigator in a tenure-track faculty position. The resulting R00 phase research aligns with NIDA Strategic Goal 2 to “develop new and improved strategies to prevent drug use and its consequences” with AI communities in the Southwest.

NIH Research Projects · FY 2026 · 2021-12

Alpha herpesviruses, including important human pathogen Herpes Simplex Virus 1 (HSV-1) and zoonotic pathogen Pseudorabies Virus (PRV), are among the very few viruses that have evolved to exploit highly-specialized neuronal cell biology. During the natural course of disease, alpha herpesviruses infect sensory and autonomic neurons of the Peripheral Nervous System (PNS). Upon reactivation, progeny virus particles undergo polarized intracellular trafficking and exocytosis at particular sub-cellular sites to spread from cell to cell, which is the subject of this proposal. In PNS neurons, this intracellular trafficking can include sorting into axons for long-distance transport into peripheral tissues, leading to recurrence of herpetic or zosteriform lesions, or to the Central Nervous System (CNS). The human alpha herpesviruses, and HSV-1 in particular, are leading causes of viral encephalitis. In Aim 1, we will investigate the viral factors that our preliminary data suggest modulate intracellular transport in non-neuronal cells, immediately prior to exocytosis, using existing reagents in PRV, and extending to HSV-1. In Aim 2, we will investigate the post-Golgi constitutive secretory pathway mechanisms that direct HSV-1 particle trafficking and exocytosis to particular sub-cellular sites in non- neuronal cells, and which we hypothesize also mediate polarized trafficking and exocytosis in neurons. Using innovative methods to acutely perturb particular cellular factors and directly image virus particle exocytosis, we will determine the role of secretory pathway mechanisms in intracellular trafficking and egress of virus particles, comparing non-neuronal cells to primary PNS neurons. In Aim 3, we will focus on the microtubule motor-based mechanisms that mediate axonal sorting, specifically in PNS neurons. Using a microfluidics-like chambered neuronal culture system and live-cell imaging, we will determine the roles of different kinesin motors and microtubule-associated proteins in axonal sorting of HSV-1 particles. Elucidating the basic cell biological processes that our viruses use in both neurons and non-neuronal cells will increase our understanding of how and why herpesviruses spread to and within the nervous system, lead to the identification of druggable targets and development of better therapies for viral neuropathology, and may provide fundamental insights into cell biology, particularly of the cell biology of neurons.

NIH Research Projects · FY 2024 · 2021-09

The proposed research is relevant to public health and well aligned with the mission of NIAMS because it addresses the substantial need to improve effectiveness in prevention of ankle sprains, one of the most common musculoskeletal injuries that can significantly impact physical activity and quality of life. While external passive supports, such as ankle taping and bracing, are the most widely used approaches to prevent ankle sprains, their long-term use makes patients overly reliant on the passive and constant support, which leads to ankle muscle and soft tissue atrophy. Furthermore, this approach not only restricts inversion-eversion motion but also dorsiflexion-plantarflexion motion, which in turn may alter natural lower-extremity biomechanics. This project seeks to study a unique smart shoe system, using integrated scientific solutions of engineering, computing and behavioral science, to overcome these limitations and provide enhanced support for individuals at risk of ankle injury. Our specific aims are: (1) to design and implement a smart shoe system that integrates robust, real-time estimation of biomechanical data and activity recognition from wearable sensors with soft actuators capable of actively adjusting the stiffness of the ankle brace; (2) to quantify and model ankle stiffness and foot loading trajectory in healthy individuals as well as in people at risk of ankle sprains; (3) to integrate an injury prediction algorithm and a closed-loop feedback controller that provides active ankle support to prevent sprains; and (4) to refine the developed algorithms for free-living use and design an intuitive mobile user interface that provides summative injury risk metrics in real-time. We will validate and evaluate the proposed smart shoe system by closely engaging with clinical partners to ensure the system is tailored toward optimal clinical integration in mind. The smart shoe system is clinically significant because it will allow clinicians to better understand foot-ankle mechanics and their correlation to ankle injury risk during various physical activities and help them provide effective and summative feedback to the users in a timely manner to promote their adherence to ankle injury prevention strategies. Successful development of the system and rigorous assessment of its patient- and clinic-centered perceived utility will lay the groundwork for follow-up clinical trials specifically aimed at investigating the long-term effect of interventions on preserving/promoting ankle health and enhancing behavioral adherence to clinical recommendations.

NIH Research Projects · FY 2024 · 2021-09

PROJECT SUMMARY/ABSTRACT: Clinical islet transplantation is a promising treatment for insulin-dependent diabetic patients, with the potential to eliminate long-term secondary complications by restoring native insulin signaling. While clinical successes have demonstrated the feasibility of achieving insulin independence through islet replacement therapy, the necessity of a long term immunosuppressive regimen limits the widespread applicability of this procedure, as the substantial risk associated with chronic immunosuppression outweighs the risk of diabetes associated morbidities. As a result, much research has explored the development of macroencapsulation devices to isolate transplanted cells from the recipient immune system. To date, these devices demonstrate limited clinical efficacy, due in large part to limited oxygen delivery to encapsulated cells. In previous work, we demonstrated the use of vasculogenic degradable hydrogels to enhance vascularization, and therefore oxygenation, at the surface of macroencapsulation devices. Despite improved vascularization, non-ideal device geometry limits encapsulated cell viability and function in vivo, as indicated by in silico modeling of device oxygenation. As such, we seek to approach macroencapsulation device design using computational modeling to optimize device oxygen distribution prior to fabrication and testing, and evaluate device oxygenation in vitro and in vivo via a novel, siloxane probe-based magnetic resonance (MR) oximetry technique, originally developed by co-PI Dr. Vikram Kodibagkar for cancer applications. We hypothesize that MR oximetry, via siloxane core probe device labelling, will enable the first precise tracking and evaluation of macroencapsulation device oxygenation in a spatiotemporal manner. We anticipate that MR imaging will validate in silico finite element modeling predictions of oxygen distribution within varied macroencapsulation device designs, and enable non-invasive, real-time tracking of macroencapsulation device oxygenation levels in vivo. These hypotheses will be addressed in the experiments of the following Specific Aims: (1) to validate in silico-optimized macroencapsulation device oxygen gradients via MR oximetry in vitro; (2) to use non-invasive MR oximetry to evaluate in vivo oxygenation of macroencapsulated cell grafts in real time; and (3) use MR oximetry to evaluate macroencapsulation devices scaled to a larger rodent model. We anticipate that this study will enable the design of improved macroencapsulation devices that significantly enhance encapsulated cell survival and function in vivo. This approach to device design, validation, and in vivo evaluation may also facilitate the process of device scale-up, potentially streamlining the process of macroencapsulation device translation to the clinic.

NIH Research Projects · FY 2025 · 2021-09

Summary One of the critical challenges in the treatment of Glioblastoma (GBM) is the presence of highly resistant cells with stem-like properties, called glioma stem cells (GSCs), that evade surgical resection, resist conventional treatments and are primarily responsible for tumor recurrence. The perivascular niche within the GBM tumor microenvironment (TME) has been well recognized as a critical site that shelters GSCs and promotes their stemness, invasion, and therapeutic resistance. Extensive studies from others and our lab, using in vitro and in vivo models, have demonstrated that the crosstalk between the endothelial cells (ECs) and GSCs regulates GSC proliferation, tumorigenicity and self- renewal capacity. However, the perivascular niche is a complex microenvironment comprised not only of ECs but multiple other cell types including astrocytes, pericytes, and immune cells. How the cell-cell interactions between the various cellular components of the perivascular niche modulate GSC behavior (proliferation vs. quiescence and invasion vs. homing) and therapy resistance is poorly understood. To address these unmet biological knowledge gaps, there is a critical need for sophisticated and more realistic ex vivo tumor models that better recapitulate the physiological complexities of the GBM perivascular niche to advance our fundamental understanding of the biology of the disease and predict therapeutic responses. Recently, we have established and validated an on-chip microfluidic tumor model of GBM, with a unique 3D organotypic architecture, to study the influence of the perivascular niche on GSC invasion. We have shown that co-culturing of astrocytes enhances EC-induced invasion of GSCs, where RNA-seq analysis of mono-culture vs. tri-culture provided a mechanistic insight into the receptor-ligand pairs that mediate the interactions between cells. Based on these foundational developments, in this study our goal is to develop an ex vivo tumor model of GBM, bioinspired from the native perivascular niche, with patient-derived cells to dissect the role of cellular components within the niche on GSC biology and response to treatment at single cell resolution. In Aim 1, our objective is to determine the influence of the key cell types within the perivascular niche on GSC-EC interactions. In Aim 2, we plan to mechanistically unveil the impact of radiation treatment on GSCs- perivascular niche interactions, while in Aim 3, we will blunt invasion and sensitize GSCs through disruption of niche-tumor cell interactions. Our study design uniquely employs an interdisciplinary approach including microengineering of a bioinspired ex vivo tumor model, single-cell level resolution analysis, molecular-level transcriptomics, and validation using ex vivo patient tumor samples. Successful completion of these studies will not only further our understanding of the interactions of GSCs with the perivascular niche but will also facilitate identification of novel targets to block disease progression.

NIH Research Projects · FY 2024 · 2021-09

Project Summary/Abstract The ultimate goal in clinical transplantation is robust and specific host immunological tolerance toward transplanted allogeneic tissue and cells. While systemic immunosuppression regimens have saved lives by enabling long-term allogeneic graft survival, they come with a host of potentially severe side effects, including infections and malignancies. The development of a method to induce donor-specific, localized tolerance toward allogeneic grafts, and thereby eliminate the need for chronic systemic immunosuppression, would dramatically reduce the risks associated with cell and tissue transplantation. Typical tolerogenic immunomodulatory approaches are systemic, potentially resulting in off-target and detrimental effects. Many groups have turned toward local delivery or presentation of immunomodulatory factors, which can eliminate systemic and off-target effects, but have limited opportunity for renewal when exhausted, resulting in a transient treatment. One approach to achieve more persistent localized immunomodulation includes the use of tolerogenic cells, such as T regulatory cells, tolerized mesenchymal stem cells or tolerized antigen presenting cells. However, while these cells can be influenced to take on a tolerogenic phenotype in vitro, it is unclear whether, or for how long, these manipulated cells maintain this phenotype in vivo. An alternative strategy would be the use of professional tolerogenic cells, such as placenta-derived trophoblasts, wherein their primary physiological role is sustained and persistent maintenance of tolerance toward allogeneic tissue. Our research approaches immunological tolerance from the perspective of the only physiologically natural scenario of allogeneic tissue tolerance, placental pregnancy. In placental pregnancy, semi-allogeneic and fully allogeneic conceptus are protected from immune response by the placenta, a fetus-derived organ consisting of various types of trophoblasts which physically isolate the fetus from the mother. We aim to probe and exploit the tolerogenic immunomodulatory mechanisms of trophoblasts and their capacity to induce tolerance in allogeneic graft transplantation through three main projects: (1) develop artificial placenta organoids using tunable hydrogel matrices to direct trophoblast differentiation and explore their immunomodulatory mechanisms in vitro; (2) engineer a safe and translatable method of artificial placenta organoid transplantation for tolerogenic cell therapy; (3) develop a tolerance-inducing vaccine via implantation of artificial placenta organoids for antigen-specific tolerance against allogeneic grafts. The use of placenta-based mechanisms to induce tolerance toward allogeneic grafts is a drastically understudied area of immunology, and presents an opportunity to widen and enrich the broader research areas of transplantation and transplantation immunology.